CN103792204A - Rapid microbiological detection technology based on terahertz time-domain spectroscopy - Google Patents
Rapid microbiological detection technology based on terahertz time-domain spectroscopy Download PDFInfo
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Abstract
The invention relates to a rapid microbiological detection technology based on terahertz time-domain spectroscopy. The rapid microbiological detection technology comprises the following steps: (1) separating and purifying microbes, and preparing microbiological suspensions with different concentration gradients; (2) scanning cell suspensions of different microbes by utilizing a terahertz time-domain spectrometer, and establishing a database for a scanning curve of different microbes; (3) performing single wavelength detection on the cell suspensions with different concentrations of the same microbe, and establishing a mathematical simulation relationship between the concentration and a detection value; (4) preparing a bacterium suspension from the sample according to a microbiological method; (5) calculating the number of various microbes in the sample according to the terahertz time-domain spectrum of the sample bacterium suspension subjected to column chromatography isolation under single frequency and the mathematical simulation relationship in the curve (3); and (6) performing full-wavelength scanning on the microbiological suspension under the same absorption peak, comparing a scanning result with the database in the step (2), thus determining the types of microbes.
Description
Technical field:
The invention belongs to biological technical field, be specifically related to a kind of microbial rapid detection technology based on terahertz time-domain spectroscopic technology.
Background technology:
Food-safety problem is related to human health and the international people's livelihood, food of a great variety, and, in various food, all may there is the inhibition factor that hinders detection accuracy in complicated component, and current various method for quick are a just reference in practice also.Microorganism specific enzyme rapid reaction detection technique detection method sensitivity viral in conventional food is too low, and technique of analytical chemistry, carrier technique, metabolism technology, immunoassay detection technique have improved the sensitivity and the selectivity that detect greatly; The domestic and international context of detection at food-borne virus, in conventional food, the detection method of virus has electron microscopic observation, cell cultivation, nucleic acid hybridization, enzyme linked immunological and polymerase chain reaction, but the sensitivity of electron microscopic observation, nucleic acid hybridization and enzyme-linked immune detection method is relatively all very low, can't be applied to separately detection; The operation of cell culture method is loaded down with trivial details, take longlyer, generally need just can observe for one week cell pathology reaction, the more important thing is that many enteron aisle food-borne virus can not carry out cell cultivation, or can cultivate but not occur cytopathic effect, this has brought very large difficulty and challenge to testing.The molecular biology method of development is that virus detection quick, sensitive in food has brought hope now, and because most food-borne virus is RNA virus, so reverse transcription PCR (RT-PCR) is applied more, but, there is false positive and Problem of False Negative in the elution of virus, concentrated and nucleic acid extraction flow process complexity.
How to set up fast, accurately, trace, multiobject detection method more and more receive researcher's concern.Not only food security aspect needs the Fast Detection Technique of microorganism, and quick, the accurate detection technique of microorganism is also badly in need of in other fields such as biology and medical science.This technology utilizes terahertz time-domain spectroscopic technology multiple-microorganism to be separated and realized qualitative and quantitative analysis according to the different chromatographic resolution principles of using for reference of the Terahertz scanning curve of different microorganisms, overcome the shortcoming of molecular biology method, simplify trace routine, make the detection method of microorganism to robotization, rapid, highly sensitive, high specificity, reproducible, and simple and easy easy row; Test condition is easy to high precision and the high sensitivity of standardization and detection.Along with the development of economic society, will inevitably more and more higher requirement be proposed to the fast detecting of microorganism, along with the continuous progress of each subject research, also there will be more simple and quick sex pheromone detection method and detection system simultaneously.
Summary of the invention:
The object of this invention is to provide a kind of microbial rapid detection technology based on terahertz time-domain spectroscopic technology, this technology utilizes advanced terahertz light analysis of spectrum to realize the fast quantification qualitative analysis to microorganism.It is worth mentioning that all belong to this patent interest field in similar to this technical method thinking.
For achieving the above object, the present invention is by the following technical solutions:
(1) microorganism is carried out to separation and purification, and prepare the microbial suspension of variable concentrations gradient; (2) utilize terahertz time-domain spectroscopy instrument to scan the cell suspending liquid of different microorganisms, by the scanning curve building database (as shown in Figure 1) of different microorganisms; (3) utilize the cell suspending liquid of the variable concentrations of terahertz time-domain spectroscopy instrument to same microorganism to carry out single wavelength detection, and concentration and detected value are set up to mathematical simulation relation; (4) sample is prepared bacteria suspension according to micro-biological process; (5) sample bacteria suspension process terahertz time-domain spectroscopy instrument continuous detecting under single-frequency after column chromatography for separation, and detection data are depicted as to curve (as shown in Figure 2), be related to the quantity of each quasi-microorganism in calculation sample according to the mathematical simulation in curve (3); (6) microbial suspension under same absorption peak is carried out to full wavelength scanner and scanning result and the database in (2) are compared, thus the kind of definite microorganism.
In described step (1), the microorganism of separation and purification comprises the microbe species in described testing sample.
In described step (2), the Terahertz mark scanning collection of illustrative plates that building database is microorganism, its scanning wavelength is that Terahertz all-wave is long, database is for comparing with the scanning spectra of testing sample microorganism after separating.
In described step (3), selected single wavelength and sample detection consistent wavelength, single wavelength is any one wavelength in Terahertz all-wave length.
Accompanying drawing explanation
Accompanying drawing 1 is different microorganisms mark scanning figure; Accompanying drawing 2 is simulation post after separating detection curve.
Embodiment
Embodiment:
The scope of protection of present invention is not limited to the scope that embodiment represents.
Comprise the following steps: (1) is carried out separation and purification, and prepared respectively 10 Escherichia coli, staphylococcus aureus, three kinds of microorganisms of bacillus subtilis
1～10
12microbial suspension under concentration gradient; (2) utilize terahertz time-domain spectroscopy instrument to carry out full wavelength scanner to the cell suspending liquid of three kinds of microorganisms, and collection of illustrative plates is saved in database; (3) utilize terahertz time-domain spectroscopy instrument respectively the cell suspending liquid of the variable concentrations to three kinds of microorganisms carrying out single wavelength detection, and respectively each microorganism concn and its detected value are set up to mathematical simulation relation; (4) sample is prepared bacteria suspension according to micro-biological process; (5) sample bacteria suspension process terahertz time-domain spectroscopy instrument continuous detecting under 5THz condition after column chromatography for separation, and detection data are depicted as to curve, be related to the quantity of each quasi-microorganism in calculation sample according to the mathematical simulation in curve (3); (6) microbial suspension under same absorption peak is carried out to full wavelength scanner and scanning result and the database in (2) are compared, thus the kind of definite microorganism.
Claims (5)
1. the microbial rapid detection technology based on terahertz time-domain spectroscopic technology, is characterized in that, comprises the following steps: the foundation of (1) database: 1. microorganism is carried out to separation and purification, and prepare the microbial suspension of variable concentrations gradient; 2. utilize terahertz time-domain spectroscopy instrument to scan the cell suspending liquid of different microorganisms, by the mark scanning graphs building database (as shown in Figure 1) of different microorganisms; 3. utilize the cell suspending liquid of the variable concentrations of terahertz time-domain spectroscopy instrument to same microorganism to carry out single wavelength detection, and concentration and detected value are set up to mathematical simulation relation.(2) sample detection: 1. sample is prepared bacteria suspension according to micro-biological process; 2. sample bacteria suspension, through terahertz time-domain spectroscopy instrument continuous detecting after column chromatography for separation, and is depicted as curve (as shown in Figure 2) by detection data, and the mathematical simulation according to Database step in is 3. related to the micro organism quantity of calculation sample; 4. the microbial suspension under same absorption peak is carried out to length scanning and by scanning result and Database step the database in 2. compare, thereby determine the kind of microorganism.
2. method according to claim 1, is characterized in that: the microorganism of separation and purification comprises the microbe species in described testing sample.
3. according to method claimed in claim 1, it is characterized in that: the bacteria suspension concentration of preparation is 10 ~ 10
12cfu/mL.
4. according to the method described in claim 2, it is characterized in that: the Terahertz mark scanning collection of illustrative plates that building database is microorganism, its scanning wavelength is that Terahertz all-wave is long, database is for comparing with the scanning spectra of testing sample microorganism after separating.
5. according to the method described in claim 3, it is characterized in that: selected single wavelength and sample detection consistent wavelength, single wavelength is any one wavelength in Terahertz all-wave length.
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Cited By (8)
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| CN104152531A (en) * | 2014-08-15 | 2014-11-19 | 中国人民解放军第三军医大学第一附属医院 | Method for establishing pathogenic bacterium fingerprint by using Terahertz waves |
| CN104502303A (en) * | 2015-01-20 | 2015-04-08 | 中国人民解放军第三军医大学第一附属医院 | Sub-THz nano-biosensor for quickly frame-detecting bacteria and detection method thereof |
| CN104764715A (en) * | 2015-04-24 | 2015-07-08 | 南京大学 | Manufacturing method for novel multi-frequency-point high-sensitivity terahertz sensor |
| CN105004671A (en) * | 2015-07-07 | 2015-10-28 | 中国人民解放军第三军医大学第一附属医院 | Water content-based terahertz spectrum detection cell for rapid and label-free detection of pathogenic bacteria, and method thereof |
| CN105486625A (en) * | 2016-01-28 | 2016-04-13 | 中国科学院重庆绿色智能技术研究院 | Cell counting device and method based on Terahertz time-domain spectroscopy technology |
| CN106404709A (en) * | 2016-11-21 | 2017-02-15 | 电子科技大学中山学院 | Inorganic fluorescent powder detection method based on partial least squares regression model |
| CN111366556A (en) * | 2020-04-29 | 2020-07-03 | 蓝科微电子(深圳)有限公司 | Terahertz detection method and system for microorganisms and organism inclusion |
| CN113916826A (en) * | 2021-09-13 | 2022-01-11 | 东莞理工学院 | Aviation fuel oil microbial pollution detection method based on terahertz time-domain spectroscopy technology |
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| CN104152531A (en) * | 2014-08-15 | 2014-11-19 | 中国人民解放军第三军医大学第一附属医院 | Method for establishing pathogenic bacterium fingerprint by using Terahertz waves |
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| CN104764715A (en) * | 2015-04-24 | 2015-07-08 | 南京大学 | Manufacturing method for novel multi-frequency-point high-sensitivity terahertz sensor |
| CN104764715B (en) * | 2015-04-24 | 2017-05-31 | 南京大学 | A kind of manufacture method of new multifrequency point, highly sensitive Terahertz sensor |
| CN105004671A (en) * | 2015-07-07 | 2015-10-28 | 中国人民解放军第三军医大学第一附属医院 | Water content-based terahertz spectrum detection cell for rapid and label-free detection of pathogenic bacteria, and method thereof |
| CN105486625A (en) * | 2016-01-28 | 2016-04-13 | 中国科学院重庆绿色智能技术研究院 | Cell counting device and method based on Terahertz time-domain spectroscopy technology |
| CN106404709A (en) * | 2016-11-21 | 2017-02-15 | 电子科技大学中山学院 | Inorganic fluorescent powder detection method based on partial least squares regression model |
| CN111366556A (en) * | 2020-04-29 | 2020-07-03 | 蓝科微电子(深圳)有限公司 | Terahertz detection method and system for microorganisms and organism inclusion |
| CN113916826A (en) * | 2021-09-13 | 2022-01-11 | 东莞理工学院 | Aviation fuel oil microbial pollution detection method based on terahertz time-domain spectroscopy technology |
| CN113916826B (en) * | 2021-09-13 | 2024-07-05 | 东莞理工学院 | Aviation fuel microbial contamination detection method based on terahertz time-domain spectroscopy technology |
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Application publication date: 20140514 |